Methods Of Treatment Of Keloid Using An Anti-VEGF Agent

ABSTRACT

Methods for treating pterygium recurrence following pterygiectomy, and for treating keloid recurrence, following surgical removal of the keloid, are disclosed. The methods include administering an anti-VEGF agent (e.g., antibody (e.g., bevacizumab) or small molecule inhibitor of VEGF signaling), or a combination therapy that includes co-administering an anti-VEGF agent, with an anti-inflammatory steroid and/or a non-steroidal anti-inflammatory drug (NSAID) to a subject.

RELATED APPLICATIONS

The present application is a continuation of U.S. application Ser. No.16/740,143, filed Jan. 10, 2020, which is a continuation of Ser. No.15/700,992, filed Sep. 11, 2017, now U.S. Pat. No. 10,556,008, which isa continuation of U.S. application Ser. No. 14/861,943, filed Sep. 22,2015, now U.S. Pat. No. 9,757,452, which is a division of U.S.application Ser. No. 14/261,198, filed Apr. 24, 2014, now U.S. Pat. No.9,161,978, which is a division of U.S. application Ser. No. 13/830,819,filed Mar. 14, 2013, now U.S. Pat. No. 8,747,852, which claims thebenefit of U.S. Provisional Patent Application Ser. No. 61/746,778,filed Dec. 28, 2012. The foregoing prior applications are hereinincorporated by reference in their entireties.

TECHNICAL FIELD

The present disclosure relates to methods of treating eye and skinconditions, in particular, methods for treating pterygium and keloid,including treating recurrence of these conditions following surgery.

BACKGROUND

Pterygium is a fibrovascular conjunctiva proliferation of thesupra-ocular surface with extension to the cornea. Surgery is theaccepted method for primary treatment of pterygium; however, one of themajor limitations of surgical management of pterygium is the highrecurrence rate. The growth factor, vascular endothelial growth factor(VEGF), has been shown to be highly expressed in pterygium and in thegrowing blood vessels that are associated with pterygium recurrence.

Bevacizumab (Avastin®) is a 149-Kd full length humanized anti-vascularendothelial growth factor (anti-VEGF) antibody that has been usedoff-label as a therapy for ocular neovascular diseases, such asage-related macular degeneration. Others have also found thatsubconjunctival administration of bevacizumab provided temporarytreatment of recurrent pterygium.

Keloid is a chronic skin condition that results in formation oftumor-like growths on the skin. Despite its benign nature, keloid cancause severe aesthetic and, in some cases, functional problems whichnegatively impacts a person's quality of life. Like pterygia, keloidtissue expresses excessive amounts of VEGF, which may play a role in theformation and evolution of keloid.

SUMMARY OF THE INVENTION

In certain aspects, the present disclosure provides a method fortreating pterygium recurrence comprising administering to an affectedeye of a subject in need of such treatment (1) an anti-VEGF agent; andone or both of (2) an anti-inflammatory steroid and (3) a non-steroidalanti-inflammatory drug (NSAID). In certain aspects, the method comprisesadministering (1) an anti-VEGF agent and (2) an anti-inflammatorysteroid to the affected eye of the subject. In certain aspects, thetreatment comprises administering (1) an anti-VEGF agent and (3) anNSAID to the affected eye of the subject. In some aspects, the treatmentcomprises administering (1) an anti-VEGF agent, (2) an anti-inflammatorysteroid and (3) an NSAID to the affected eye of the subject. In certainaspects, the anti-VEGF agent is an antibody or small molecule inhibitorof VEGF signaling. In other aspects, the anti-VEGF agent is an antibody.In some aspects, the anti-VEGF agent is bevacizumab. In certain aspects,the amount of bevacizumab administered is about 2.5 mg. In otheraspects, the anti-VEGF agent is injected at a site of new blood vesselgrowth in the affected eye. In other aspects, the anti-VEGF agent isadministered by injection. In some aspects, the anti-inflammatorysteroid is prednisolone acetate. In certain aspects, theanti-inflammatory steroid is prednisolone acetate 1%. In other aspects,the anti-inflammatory steroid is difluprednate. In certain aspects, theanti-inflammatory steroid is difluprednate 0.05%. In some aspects, theanti-inflammatory steroid is triamcinolone acetonide. In certainaspects, the amount of triamcinolone acetonide administered is 40 mg. Instill other aspects, the anti-inflammatory steroid is in the form of anophthalmic solution or suspension. In certain aspects theanti-inflammatory steroid is administered topically. In still otheraspects, the anti-inflammatory steroid is injected. In some aspects, theanti-inflammatory steroid is co-injected with the anti-VEGF agent. Inother aspects, the NSAID is selected from the group consisting ofdiclofenac, ketorolac, bromfenac and nepafenac. In certain aspects, theNSAID is in the form of an ophthalmic solution or suspension. In otheraspects, the NSAID is selected from the group consisting of diclofenac0.1%, ketorolac tromethamine ophthalmic solution 0.4%, bromfenacophthalmic solution 0.09%, and nepafenac ophthalmic suspension 0.1%. Insome aspects, the NSAID is administered topically.

In some aspects, the above treatments are performed in conjunction withsurgery to remove pterygium. In certain aspects, the treatment iscommenced within 2 weeks prior to or following the surgery to removepterygium. In other aspects, the treatment is commenced within 1 weekprior to or following the surgery to remove pterygium. In certainaspects, the treatment is commenced following surgery to removepterygium. In other aspects, the treatment is commenced within 2 weeksfollowing the surgery to remove pterygium. In other aspects, thetreatment is commenced within 1 week following the surgery to removepterygium. In certain aspects, the commencement of the treatmentcomprises administering at least one of (1) the anti-VEGF agent; (2) theanti-inflammatory steroid and (3) the non-steroidal anti-inflammatorydrug (NSAID). In certain aspects, the treatment comprises more than oneadministration of the anti-VEGF agent to the affected eye of thesubject. In other aspects, the treatment comprises more than oneadministration of the anti-inflammatory steroid or the NSAID or both tothe affected eye of the subject. In certain aspects, the anti-VEGF agentis administered at least once before the anti-inflammatory steroid orthe NSAID or both are administered. In other aspects, theanti-inflammatory steroid or the NSAID is administered to the affectedeye of the subject at least once before the anti-VEGF agent isadministered. In some aspects, the anti-inflammatory steroid and theNSAID are each administered to the affected eye of the subject at leastonce before the anti-VEGF agent is administered. In other aspects, theanti-VEGF agent, the anti-inflammatory steroid and the NSAID are eachadministered on separate days at least once. In still other aspects, theanti-VEGF agent and the anti-inflammatory steroid are administered atleast once on the same day. In still other aspects, the subject has hadpterygium recurrence prior to commencement of the treatment. In certainaspects, the subject is human.

In another aspect, the present disclosure provides a method for treatingkeloid recurrence, the method comprising administering (1) an anti-VEGFagent to a site where a keloid has been removed from a subject. Incertain aspects, the anti-VEGF agent is an antibody or small moleculeinhibitor of VEGF signaling. In other aspects, the anti-VEGF agent is anantibody. In some aspects, the anti-VEGF agent is bevacizumab. Incertain aspects, the bevacizumab is administered at a dose of 10 mg. Inother aspects, the anti-VEGF agent is administered by injection ortopically. In certain aspects, the anti-VEGF agent is injected at oradjacent to a site of new blood vessel growth at the site of keloidremoval. In some aspects, the treatment further comprises administeringto the site (2) an anti-inflammatory steroid or (3) a non-steroidalanti-inflammatory drug (NSAID), or both. In certain aspects, thetreatment further comprises administering (1) an anti-VEGF agent, (2) ananti-inflammatory steroid and (3) a non-steroidal anti-inflammatory drug(NSAID) to the site. In certain aspects, the anti-inflammatory steroidis co-administered with the anti-VEGF agent. In other aspects, theanti-inflammatory steroid is applied topically, systemically, or byinjection to the site of keloid removal. In certain aspects, theanti-inflammatory steroid is alclometasone, diflorasone, fluocinonide,or prednicarbate. In some aspects, the anti-inflammatory steroid isalclometasone 0.005% cream, diflorasone 0.005% cream, fluocinonide cream0.1%, or prednicarbate topical 0.1% cream or ointment. In certainaspects, the NSAID is diclofenac. In some aspects, the NSAID isdiclofenac 1% solution. In other aspects, the treatment furthercomprises administering to the site one or more subsequentadministrations of the anti-VEGF agent. In some aspects, the treatmentfurther comprises administering to the site one or more subsequentadministrations of one or more of an anti-VEGF agent, ananti-inflammatory steroid, and an NSAID. In certain aspects, the subjectis human.

Unless otherwise defined, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which this disclosure pertains. In case of conflict, thepresent document, including definitions provided herein, will control.

All publications, patent applications, patents, GenBank® Accessionnumbers, protocols and other references mentioned herein areincorporated by reference in their entirety for all purposes. Thematerials, methods, and examples disclosed herein are illustrative onlyand not intended to be limiting.

The details of one or more embodiments of the invention are set forth inthe accompanying drawings and the description below. Preferred methodsand materials are described below, although methods and materialssimilar or equivalent to those described herein can also be used in thepractice or testing of the present invention. Other features, objects,and advantages of the invention will be apparent from the descriptionand drawings, and from the claims.

DESCRIPTION OF DRAWINGS

FIGS. 1-5 are timelines showing the treatment regimens of Patients 1-5.

DETAILED DESCRIPTION

The present disclosure provides methods for treating pterygiumrecurrence. Pterygia are caused by a neovascular condition of the eyethat can eventually lead to blindness. In certain aspects, the methodsdisclosed herein include administering a combination therapy to anaffected eye of a subject with recurrent pterygia. First, a pterygium isremoved (e.g., surgically) from an affected eye, and then, followingremoval, the patient is treated with a combination therapy. Thecombination therapy disclosed herein includes treating the affected eyewith an anti-VEGF agent (such as, e.g., the antibody, bevacizumab, orother inhibitor of VEGF signaling) and with an anti-inflammatory steroid(e.g., triamcinolone acetonide (injected), or prednisolone acetate 1% ordifluprednate 0.05% (topical)) and/or a non-steroidal anti-inflammatorydrug (NSAID) (e.g., diclofenac 0.1%, ketorolac tromethamine ophthalmicsolution, 0.4%, bromfenac ophthalmic solution, 0.09%, nepafenacophthalmic suspension 0.1%). As disclosed herein, the anti-inflammatorysteroid and/or NSAID can be administered before or after the eye istreated with the anti-VEGF agent.

Since it has been reported in the literature that treatment of recurrentpterygia using the anti-VEGF antibody bevacizumab was not effective oronly temporarily effective (e.g., 6 months or less), it was surprisingto discover, as disclosed herein, that bevacizumab, when administered aspart of a combination therapy, was effective for inhibiting pterygiumrecurrence. For example, as shown in FIGS. 1, 2, 3A, 3B, 4, and 5,patients receiving a combination therapy including administration ofbevacizumab, steroid and NSAID, have not had recurrence of pterygium inmore than 11 months, 8 months, 6 months, 9 months, and 3 months,respectively.

The present disclosure also provides methods for treating keloidrecurrence. The methods disclosed herein include administering to thesite where a keloid has been removed an anti-VEGF agent, such as, butnot limited to an anti-VEGF antibody, e.g., bevacizumab. While notintending to be limited to any one particular theory or mechanism ofaction, keloids express high levels of VEGF, and thus the administrationof anti-VEGF antibody is believed to inhibit neovascularization, whichis required for keloid growth. The methods disclosed herein also includeadministering the anti-VEGF agent in a combination therapy with one orboth of an anti-inflammatory steroid (e.g., alclometasone 0.005% cream,diflorasone 0.005% cream, fluocinonide cream 0.1%, or prednicarbatetopical 0.1% cream or ointment) and an NSAID (e.g., diclofenac 1%solution).

Definitions

As used herein, the term “pterygium recurrence” means the reappearanceof pterygium in the eye following removal (e.g., surgically) of primarypterygium.

As used herein, the term “subject” means any mammal, and, in particular,a human, and can also be referred to, e.g., as an individual or patient.A “subject in need of treatment” pterygium or keloid recurrenceaccording to the methods disclosed herein is a subject who is at risk ofpterygium recurrence or keloid recurrence, e.g., a patient who is aboutto undergo or has recently (e.g., within about 12, 11, 10, 9, 8, 7, 6,5, 4, 3, 2 or 1 month(s), about 2 or 1 week(s) or less) undergone, or isabout to undergo (e.g., within about 1 month, about 1 week or less)surgery to remove a pterygium or keloid.

As used herein, an “anti-VEGF agent” means an inhibitor of VEGFsignaling. Anti-VEGF agents include antibodies (e.g., bevacizumab),antibody fragments (e.g., an antibody light chain (VL), an antibodyheavy chain (VH), a single chain antibody (scFv), an F(ab′)2 fragment, aFab fragment, an Fd fragment, an Fv fragment, and a single domainantibody fragment (DAb). Fragments can be obtained, e.g., via chemicalor enzymatic treatment of an intact or complete antibody or antibodychain or by recombinant means), fusion proteins, peptide, nucleic acids(e.g., siRNA, shRNA), and other small molecules, etc. that disrupt theinteraction between VEGF (VEGF-A) and its receptor (VEGFR-1/VEGFR-2).Other, non-limiting examples of anti-VEGF agents encompassed by thepresent disclosure are provided herein below.

As used herein, the term “adjacent to”, e.g., in the context ofinjecting an anti-VEGF adjacent to or near the site of new blood vesselgrowth, means proximate to (e.g., within about 0.1 mm, 0.2 mm, 0.3 mm,0.4 mm, 0.5 mm, 1 mm, 2 mm, 3 mm, 4 mm, or 5 mm from the site of bloodvessel growth).

As used herein the terms “therapeutically effective” and “effectiveamount”, used interchangeably, applied to a dose or amount refer to aquantity of a composition, compound or pharmaceutical formulation thatis sufficient to result in a desired activity upon administration to asubject in need thereof. Within the context of the present invention,the term “therapeutically effective” refers to that quantity of acomposition, compound or pharmaceutical formulation that is sufficientto reduce, eliminate or delay at least one symptom of a disease orcondition specified herein, e.g., pterygium or keloid recurrence. When acombination of active agents is administered, the effective amount ofthe combination, or individual agents, may or may not include amounts ofeach agent that would have been effective if administered individually.The dosage of the therapeutic formulation will vary, depending upon thenature of the disease or condition, the patient's medical history, thefrequency of administration, the manner of administration, the clearanceof the agent from the host, and the like. The initial dose may belarger, followed by smaller maintenance doses. The dose may beadministered, e.g., weekly, biweekly, daily, semi-weekly, etc., tomaintain an effective dosage level.

Therapeutically effective dosages in the methods described herein can bedetermined by the treating physician. For example, the physician maybegin treatment using manufacturer-recommended doses for the anti-VEGFagent, anti-inflammatory steroid and/or NSAID, and make adjustmentsbased on the physician's observations of the effect of treatment.Further guidance is provided herein and in the Examples. In addition,clinical trials can be conducted to determine the doses that areeffective to produce statistically significant treatment effects when apopulation of patients is treated.

As used herein “combination therapy” means the treatment of a subject inneed of treatment with a certain composition or drug in which thesubject is treated or given one or more other compositions or drugs forthe disease in conjunction with the first and/or in conjunction with oneor more other therapies, such as, e.g., surgery. Such combinationtherapy can be sequential therapy wherein the patient is treated firstwith one treatment modality (e.g., drug or therapy), and then the other(e.g., drug or therapy), and so on, or all drugs and/or therapies can beadministered simultaneously. In either case, these drugs and/ortherapies are said to be “co-administered.” It is to be understood that“co-administered” does not necessarily mean that the drugs and/ortherapies are administered in a combined form (i.e., they may beadministered separately or together to the same or different sites atthe same or different times).

The phrase “pharmaceutically acceptable” is employed herein to refer tothose compounds, materials, compositions and/or dosage forms which are,within the scope of sound medical judgment, suitable for use in contactwith the tissues of human beings and animals without excessive toxicity,irritation, allergic response, or other problem or complication,commensurate with a reasonable benefit/risk ratio.

As used herein, “treating” or “treatment” of a state, disorder orcondition (e.g., pterygium or keloid recurrence) includes: (1)preventing or delaying the appearance of clinical or sub-clinicalsymptoms of the state, disorder or condition developing in a mammal thatmay be afflicted with or predisposed to the state, disorder or conditionbut does not yet experience or display clinical or subclinical symptomsof the state, disorder or condition; and/or (2) inhibiting the state,disorder or condition, including arresting, reducing or delaying thedevelopment of the disease or a relapse thereof (in case of maintenancetreatment) or at least one clinical or sub-clinical symptom thereof;and/or (3) relieving the disease, i.e., causing regression of the state,disorder or condition or at least one of its clinical or sub-clinicalsymptoms; and/or (4) causing a decrease in the severity of one or moresymptoms of the disease. The benefit to a subject to be treated iseither statistically significant or at least perceptible to the patientor to the physician.

Treating can include inhibiting pterygium recurrence in a subject. Whenpterygium recurrence is inhibited, pterygium is not detectable for atleast a predetermined time frame in the affected eye of the subjectwhere primary pterygium has been removed. For example, a predeterminedtime frame may be, e.g., at least 1 month, at least 2 months, at least 3months, at least 4 months, at least 5 months, at least 6 months, atleast 7 months, at least 8 months, at least 9 months, at least 10months, at least 11 months, or at least 12 months. Preferably, thepredetermined time frame is at least 6 months.

Treating can include inhibiting keloid recurrence in a subject. Whenkeloid recurrence is inhibited, keloid is not detectable for at least apredetermined time frame in the site where primary pterygium has beenremoved. For example, a predetermined time frame may be, e.g., at least1 month, at least 2 months, at least 3 months, at least 4 months, atleast 5 months, at least 6 months, at least 7 months, at least 8 months,at least 9 months, at least 10 months, at least 11 months, or at least12 months. Preferably, the predetermined time frame is at least 6months.

Anti-VEGF Agents

The human VEGF-A gene is organized in eight exons. Alternative exonsplicing results in the generation of four main VEGF isoforms, having,respectively, 121, 165, 189, and 206 amino acids following signalsequence cleavage (VEGF121, VEGF165, VEGF189, and VEGF206). VEGF165 isbelieved to be the most physiologically relevant isoform. For a review,see Ferrara et al., Biochem. Biophys. Res. Commun., 2005, 333, 328-335.The amino acid sequences of VEGF-A are well known in the art, and due tosplice variation, the sequences are numerous. By way of non-limitingexample, the following are exemplary and non-limiting GenBank® AccessionNos. for human VEGF-A (“VEGF”) amino acid sequences: AAP86646.1,P15692.2, NP_001191313.1, NP_001165101.1, NP_001165099.1,NP_001165097.1, NP 001165095.1, NP_001020539.2, NP_003367.4,NP_001165093.1, NP_001020541.2, NP 001191314.1, NP 001165100.1, NP001165098.1, NP 001165096.1, NP_001165094.1, NP 001028928.1, NP001020540.2, NP 001020538.2, and NP_001020537.2.

There are two VEGF receptor (VEGFR) tyrosine kinases (RTKs), Flt-1,known also as VEGFR-1 and KDR, Flk-1, or VEGFR-2. There is now agreementthat VEGFR-2 is the major mediator of the mitogenic, angiogenic, andpermeability-enhancing effects of VEGF. For a detailed review of thebiological and signaling properties of the VEGFR, see Ferrara, Endocr.Rev. 2004, 25, 581-611. The amino acid sequences for the VEGFR are knownin the art. By way of non-limiting example, GenBank® accession numbersfor VEGFR-1 amino acid sequences include (but are not limited to):NP_001153503.1, NP_002010.2, NP_001153502.1, and NP_001153392.1. Theamino acid sequences for the VEGFR-2 are known in the art. By way ofnon-limiting example, GenBank® accession numbers for VEGFR-2 amino acidsequences include (but are not limited to): NP_002244.1, AAC16450.1, andNP_001153503.1.

Disclosed herein are methods for treating pterygium or keloidrecurrence. The methods include administering an anti-VEGF agent to asubject.

The present Examples demonstrate that the anti-VEGF antibody bevacizumabcan be used in the present methods. The antibody bevacizumab and itsVEGF-binding activity are reviewed in detail in Ferrara et al., Biochem.Biophys. Res. Commun., 2005, 333, 328-335. Bevacizumab may beadministered to the eye in an amount of about 2.5 mg. However suitableranges include, e.g., from about 0.5 mg to about 10 mg, about 1 mg toabout 5 mg, about 2 mg to about 4 mg, and about 2 mg to about 3 mg. Forexample, the antibody can be administered at a dosage in the range ofabout 0.5 mg/0.1 mL to 10 mg/0.1 mL in a total volume of 0.1 mL.Bevacizumab may be administered to skin (e.g., for the inhibition ofkeloid recurrence) at a dose about 5-15 mg. In one embodiment, the dosefor administration to site of keloid removal (e.g., for inhibition ofkeloid recurrence) is about 10 mg).

It is to be appreciated, however, that the treatment method describedherein can also be performed using other anti-VEGF agents (e.g., VEGF orVEGFR inhibitors, such as, but not limited to, other anti-VEGFantibodies, drugs, prodrugs, small molecules, peptides, nucleic acidinhibitors (e.g., siRNA, shRNA, antisense oligonucleotides), fusionproteins, etc.), either that are known in the art or that will bediscovered or engineered in the future, so long as the anti-VEGF agenthas the ability to inhibit the action of VEGF (e.g., human VEGF) and/ora VEGFR (e.g., VEGFR-1 and/or VEGFR-2) (e.g., human VEGFR-1 or humanVEGFR-2) (i.e., to inhibit VEGF signaling). Assays for determiningwhether an antibody or other agent interferes with VEGF signaling(either by inhibiting VEGF or a VEGFR or the interaction between VEGFand its receptor), for example, are well known in the art, and can beused to determine whether an anti-VEGF agent interferes with VEGFsignaling and is therefore encompassed by the presently disclosedmethods. Non-limiting examples of such assays include the VEGFinhibition assays described in Vicari et al., J. Biol. Chem., 2011,286(15), 13612-25 and Brekken et al. Cancer Res., 2000, 60, 5117-24.

By way of non-limiting example, other anti-VEGF antibodies andinhibitors that are known in the art, and, that can be used in themethods disclosed herein include but are not limited to: ranibizumab,pegaptanib, imatinib, vandetanib, sorafenib, pazopanib, valatanib,vevasiranib, aflibercept, etanercept, anecortave acetate (angiostaticsteroid), VEGF-trap (a fusion protein), squalamine lactate, erlotinib,gefitinib (small molecules), Combretastatin A4 Prodrug (anantitubulin/antiangiogenic agent), AdPEDF (Adenovector pigmentepithelium-derived factor), Cand5 (siRNA), protein tyrosine kinase 7inhibitors (PTK7), lipolytic agents, TG100801, AG013958, AL39324,AGN211745 (VEGF receptor blockers), anti-angiogenic VEGF-A(xxx)b family,VEGF Trap (receptor decoy), protein kinase antibodies to tyrosine kinaseinhibitor receptors SIM010603, kinase domain receptor antibodies (KDR1.3and KDR2.6), GS101 aganirsen (an antisense oligonucleotide againstinsulin receptor substrate aka IRS-1), picropodophyllin (PPP),tetrameric tripeptide, tissue kallikrein, KH906 (a recombinant humanVEGF receptor protein fusion), beta-adreno receptor blocker β3-AR,nicotinic acetycholine receptor antagonists, linomide analogue (Lin05),morpholino oligomers (VEGFR1_MOe13), decursin, prorenin, vasohibin andsirolimus. It will be appreciated that because the amino acids sequences(as well as nucleic acid sequences encoding the amino acid sequences) ofVEGF and VEGFRs are known in the art, the skilled artisan can readilydesign additional anti-VEGF agents for use in the presently disclosedmethods.

Dosage ranges for anti-VEGF agents, e.g., those disclosed above, can bereadily determined by the ordinarily skilled artisan, and can, e.g.,first be determined in animal models for determining dosage, safety andefficacy according to standard methods known in the art.

Anti-Inflammatory Steroids

Anti-inflammatory steroids are steroidal compounds that haveanti-inflammatory activity and include corticosteroids, includingglucocorticoids. Glucocorticoids bind to glucocorticoid receptors in thecytoplasm which may increase the transcription of genes coding foranti-inflammatory proteins, including lipocortin-1, interleukin-10,interleukin-1 receptor antagonist and neutral endopeptidase.Glucocorticoids also inhibit the expression of multiple inflammatorygenes, including genes for various cytokines, enzymes, receptors andadhesion molecules. Barnes et al., Clin. Sci., 1998, 94, 557-572.

Suitable anti-inflammatory steroids for injection into the eye andsuitable for use in the methods described herein include, e.g.,triamcinolone acetonide (Kenalog®, e.g., Kenalog®-40, available fromRanbaxy Laboratories Inc. (0.0147% spray), Triesence® available fromAlcon Laboratories Inc., Trivaris® available from Allergan Inc.,Trianex® available from Upsher Smith Laboratories Inc.). Suitabledosages of triamcinolone acetonide range from 10 mg/mL to 40 mg/mL withvolume given range from 0.5 mL to 1 mL. These steroids can beco-injected with other agents described herein (e.g., with the anti-VEGFantibody). Further injectable anti-inflammatory steroids suitable foruse in the methods described herein are listed in Table 1.

TABLE 1 Commercially Available Injectable Steroids Steroid TradenamesMethylprednisolone Depo-Medrol ®; Solu-Medrol ® DexamethasoneDecadron ®-LA; Decadron ® Phosphate Betamethasone Celestone ®Hydrocortisone Various Triamcinolone Aristocort ®, Kenalog ®

Suitable steroids for topical administration to the eye (e.g., as eyedrops or ointment) include, e.g., prednisolone acetate 1% (e.g., PredForte®, Allergan Inc.) or difluprednate 0.05%. Suitable dosages foradministration to humans include, e.g., one drop to eye with frequencyranging from every hour to every 24 hours depending on the degree ofinflammation characterized by redness, swelling, pain and intraocularpressures. Further commercially available topical ocular steroidssuitable for use in the methods described herein are listed in Table 2.

TABLE 2 Commercially Available Topical Ocular Steroids Concentration/Steroid Tradenames Formulation Prednisolone acetate Pred Forte ® 1.0%suspension Econopred ® 1.0% suspension Plus (Alcon) 1.0% suspensionAK-Tate ™ 0.125% suspension Pred Mild ® 0.125% suspension Econopred ®Prednisolone Inflamase ® 1.0% solution sodium Forte 0.5% solutionphosphate Metreton ™ 0.125% solution Inflamase ® 0.125% solution MildAK-Pred ™ Dexamethasone Maxidex ® 0.1% suspension alcohol Tobradex ®ointment 0.1% suspension ointment Fluorometholone Flarex ® 0.1%suspension acetate Eflone ™ 0.1% suspension Fluorometholone FML ® 0.1%suspension alcohol Fluor-Op ™ 0.1% ointment FML ®-Mild 0.25% suspensionRimexolone Vexol ® 1% suspension Medrysone HMS ® 1.0% suspension alcoholLotoprednol Lotemax ® 0.5% suspension etabonate Alrex ® 0.2% suspensionZylet ® 0.5% suspension

Suitable steroids for application to the skin include, e.g.,alclometasone 0.050% cream (generic or Aclovate® 0.050% cream availablefrom PharmaDerm Inc.), diflorasone 0.0050% cream, prednicarbate 0.100(generic, or Dermatop 0.100 cream or ointment available fromSanofi-Aventis US LLC), and fluocinonide cream 0.1% (e.g., Vanos®,available from Medicis Inc.). Suitable dosages for administration tohumans include, e.g., 0.05% to 0.1%. Further commercially availabletopical ocular steroids suitable for use in the methods described hereinare listed in Table 3.

TABLE 3 Commercially Available Topical Steroids Potency Drug Super-HighBetamethasone dipropionate, augmented Clobetasol propionate FluocinonideFlurandrenolide Halobetasol propionate High Amcinonide Betamethasonedipropionate Desoximetasone Diflorasone diacetate FluocinonideHalocinonide Triamcinolone Medium-High Amcinonide Betamethasonedipropionate Betamethasone valerate Desoximetasone Diflorasone diacetateFluocinonide Fluticasone propionate Mometasone furoate Triamcinoloneacetonide Medium Clocortolone pivalate Fluocinolone acetonideHydrocortisone valerate Mometasone furoate Triamcinolone acetonideMedium-Low Betamethasone valerate Desonide Fluocinolone acetonideFluticasone proprionate Hydrocortisone butyrate Hydrocortisone probutateHydrocortisone valerate Prednicarbate Triamcinolone acetonide LowAclometasone dipropionate Betamethasone valerate Desonide Fluocinoloneacetonide Triamcinolone acetonide Very Low Hydrocortisone (base)Hydrocortisone acetate/Aloe vera Hydrocortisone acetate/urea

Non-Steroidal Anti-Inflammatory Drugs (NSAIDs)

Non-steroidal anti-inflammatory drugs (NSAIDs) are non-steroidalcompounds that reduce inflammation. Most NSAIDs act as nonselectiveinhibitors of the enzyme cyclooxygenase (COX), inhibiting both thecyclooxygenase-1 (COX-1) and cyclooxygenase-2 (COX-2) isoenzymes. COXcatalyzes the formation of prostaglandins. Since COX-1 inhibition isbelieved to be associated with gastrointestinal side-effects of NSAIDs,compounds that are selective COX-2 inhibitors have also been developed.

Non-limiting examples of NSAIDs that can be administered to the eyeinclude, e.g., diclofenac 0.1%, ketorolac tromethamine ophthalmicsolution, 0.4% (Allergan Inc.) bromfenac ophthalmic solution, 0.09%(Bausch and Lomb Inc.), nepafenac ophthalmic suspension 0.1% (AlconLaboratories Inc.) fluriprofen sodium 0.03% solution and ketorolactromethamine 0.5% solution. Suitable dosage regimens for administrationto humans include, e.g., 0.1% applied one drop once a day, up to 4 timesa day (every 6 hours).

Non-limiting examples of NSAIDs that can be administered to the skininclude, e.g., diclofenac 1% (topical) (e.g., Voltaren gel 1% fromNovartis Pharmaceuticals Corp. or Pennsaid 1.5% solution (topical) fromMallinckrodt Pharmaceuticals). Suitable dosages for administration tohumans include, e.g., 1% (topical) apply to skin once a day to 40 mg(administer by injecting 1 mL into skin).

Methods of Treating Pterygium and Keloid Recurrence

Provided herein are methods for treating (e.g., inhibiting) pterygiumrecurrence in an affected eye of a subject, the method including (e.g.,comprising, consisting essentially of, consisting of) administering tothe affected eye of a subject in need of such treatment (1) at least oneanti-VEGF agent (e.g., ranibizumab, pegaptanib, anecortave acetate(angiostatic steroid), VEGF-trap (a fusion protein), squalamine lactate,erlotinib, gefitinib (small molecules), combretastatin A4 prodrug (anantitubulin/antiangiogenic agent), AdPEDF (Adenovector pigmentepithelium-derived factor), Cand5 (siRNA), and TG100801 (VEGF receptorblocker); and one or both of (2) an anti-inflammatory steroid (e.g.,triamcinolone acetonide, prednisolone acetate 1%, or difluprednate0.05%) and (3) a non-steroidal anti-inflammatory drug (NSAID) (e.g.,diclofenac 0.1%, ketorolac tromethamine ophthalmic solution 0.4%,bromfenac ophthalmic solution 0.09%, and nepafenac ophthalmic suspension0.1%).

In a specific embodiment, a method for treating pterygium recurrence inan affected eye of a subject includes (e.g., comprises, consistsessentially of, consists of) administering to the affected eye of asubject in need of such treatment (1) bevacizumab and (2) triamcinoloneacetonide (or prednisolone acetate 1%, or difluprednate 0.05%) and/or(3) diclofenac 0.1% (or ketorolac tromethamine ophthalmic solution 0.4%,bromfenac ophthalmic solution 0.09%, or nepafenac ophthalmic suspension0.1%).

In another specific embodiment, a method for treating pterygiumrecurrence in an affected eye of a subject includes (e.g., comprises,consists essentially of, consists of) administering to the affected eyeof a subject in need of such treatment (1) bevacizumab; and (2)triamcinolone acetonide (or prednisolone acetate 1%, or difluprednate0.05%).

In another specific embodiment, a method for treating pterygiumrecurrence in an affected eye of a subject includes (e.g., comprises,consists essentially of, consists of) administering to the affected eyeof a subject in need of such treatment (1) bevacizumab; and (2)diclofenac 0.1% (or ketorolac tromethamine ophthalmic solution 0.4%,bromfenac ophthalmic solution 0.09%, or nepafenac ophthalmic suspension0.1%).

In yet another specific embodiment, a method for treating pterygiumrecurrence in an affected eye of a subject includes (e.g., comprises,consists essentially of, consists of) administering to the affected eyeof a subject in need of such treatment (1) bevacizumab; (2)triamcinolone acetonide (or prednisolone acetate 1%, or difluprednate0.05%); and (3) diclofenac 0.1% (or ketorolac tromethamine ophthalmicsolution 0.4%, bromfenac ophthalmic solution 0.09%, or nepafenacophthalmic suspension 0.1%).

Also provided herein are methods for treating (e.g., inhibiting) keloidrecurrence, wherein the method includes (e.g., comprises, consistsessentially of, or consists of) administering to a site where a keloidhas been removed in a subject (1) an anti-VEGF agent (e.g., Ranibizumab,pegaptanib, Anecortave acetate (angiostatic steroid), VEGF-trap (afusion protein), squalamine lactate, erlotinib, gefitinib (smallmolecules), combretastatin A4 prodrug (an antitubulin/antiangiogenicagent), AdPEDF (Adenovector pigment epithelium-derived factor), Cand5(siRNA), and TG100801 (VEGF receptor blocker). In certain aspects, themethod further includes administering to the site one or both of (2) ananti-inflammatory steroid (e.g., alclometasone 0.005% cream, diflorasone0.005% cream, fluocinonide cream 0.1%, or prednicarbate topical 0.1%cream or ointment); and (3) an NSAID (e.g., diclofenac 1% solution).

In a specific embodiment, a method for treating keloid recurrence isprovided, wherein the method includes administering (1) bevacizumab. Incertain embodiments, the therapy further includes administering to thesite (2) alclometasone 0.005% cream, diflorasone 0.005% cream,fluocinonide cream 0.1%, or prednicarbate topical 0.1% cream orointment. In another embodiment, a method for treating keloid recurrenceis provided, wherein the method includes administering to the site (ofprevious keloid removal) (1) bevacizumab and (2) an NSAID diclofenacsolution or gel (e.g., 1% solution or gel). In another embodiment, amethod for treating keloid recurrence is provided, wherein the methodincludes administering to the site (of previous keloid removal) (1)bevacizumab, (2) alclometasone or diflorasone cream (e.g., 0.005%cream), or prednicarbate topical 0.1% cream or ointment; and (3) anNSAID (e.g., diclofenac 1% solution).

When a combination therapy is administered to a subject, it will beappreciated that the agents administered as part of the combinationtherapy can be administered in any order, e.g., as determinedappropriate, e.g., by the subject's physician. For example, a subjectmay be first administered an anti-inflammatory steroid (e.g., eye dropor skin cream), followed by injection or other suitable route ofadministration of an anti-VEGF agent (e.g., as disclosed herein),followed by topical administration of an NSAID (e.g., eye drop,ointment, or skin cream). However, in other embodiments, the subject maybe first administered an NSAID, followed by injection or other suitableroute of administration of an anti-VEGF agent (e.g., as disclosedherein) alone, followed by topical administration of ananti-inflammatory steroid, or followed by injection (e.g., co-injection)or other suitable route(s) of administration of the anti-VEGF agent andan injectable steroid. In another embodiment, the subject can be firstadministered an anti-VEGF agent, by a suitable route of administration,followed by an NSAID and then an anti-inflammatory steroid, or thesubject can be first administered an anti-VEGF agent by a suitable routeof administration, followed by an anti-inflammatory steroid, followed byan NSAID. In other embodiments, all two or all three agents (anti-VEGFagent, anti-inflammatory steroid and/or NSAID) can be administeredtogether, e.g., at or near the same time, e.g., within 1, 2, 3, 4, 6, 8,10, 12 or 24 hours of each other. It is also contemplated that more thanone anti-VEGF agent can be administered in combination in the methodsdisclosed herein (either at the same or different times, by the same ordifferent route(s) of administration).

The length of time between removal of the pterygium or keloid and thefirst treatment can vary according to each patient's needs. Forpterygia, it is typical to wait at least a day or two following thesurgery, for example, to inject anti-VEGF antibody, simply to give thepatient time to recover. However, it is also possible to inject theanti-VEGF antibody and/or injectable steroid, as well apply topicalsteroid and/or NSAID on the day of surgery. Treatment, e.g., with acombination therapy disclosed herein can also begin, about 2 or more,about 3 or more, about 4 or more, about 5 or more, about 6 or more, orabout 7 or more days after surgery, about 1 or 2 weeks or more, about 3weeks or more, about 4 weeks or more, or about 5 weeks or more after thesurgery. The treatment can also be commenced in preparation for surgery,e.g., by administering one or more of the drugs within about 1 weekprior to the surgery.

Time between administrations of the different components (antibody,steroid, and/or NSAID) of the combination therapy can also range widelydepending on each patient's needs. When two or more agents of thecombination therapy are not administered on the same day, they can beadministered about 1 or more, about 2 or more, about 3 or more, about 4or more, about 5 or more, about 6 or more, or about 7 or more daysapart, about 1 or 2 weeks or more, about 3 weeks or more, about 4 weeksor more, or about 5 weeks or more apart.

The methods disclosed herein provide an improved treatment of pterygiumand keloid. As discussed above, currently available treatments typicallyresult in recurrence after only a short period of time (e.g., less thanabout 6 months). The presently disclosed methods provide long-termtreatments (including complete inhibition) of pterygium and keloidrecurrence (e.g., greater than about 6, 7, 8, 9, 10 or 11 monthsfollowing removal of the pterygium or keloid). Typically, although notnecessarily, treatment with the combination therapy disclosed herein cancontinue until a patient is determined to have no new blood vesselgrowth and no pterygium recurrence for about 6 months from the time ofthe last treatment. At that time, the patient may continue to receiveNSAID drug (e.g., NSAID eye drops), which may be administered for up to,e.g., about one year.

Administration

Compositions and formulations including an antibody, steroid and/orNSAID (“agent”) as described herein can be administered topically orparenterally, or by any other suitable methods known in the art. Theterm “parenteral” includes injection or deposition or sustained releasevia vehicles or devices (e.g., intravenous, subconjunctival, subtenon,episcleral, intrascleral, subscleral, intraperitoneal, epidural,intrathecal, intramuscular, intraluminal, intratracheal, epidermal,intradermal, subdermal or subcutaneous). Moreover, the different agentsadministered in the combination therapy disclosed herein may beadministered by different routes. For example, an antibody or otheranti-VEGF agent disclosed herein may be injection into the eye or skin,or applied topically; an anti-inflammatory steroid and/or or NSAID maybe administered systemically (e.g., by injection), orally, and/ortopically, e.g., to the eye or skin.

While it is possible to use an agent disclosed herein for therapy as is,it may be preferable to administer the agent as a pharmaceuticalformulation, e.g., in admixture with a suitable pharmaceuticalexcipient, diluent, or carrier selected with regard to the intendedroute of administration and standard pharmaceutical practice.Pharmaceutical formulations include at least one active compound, inassociation with a pharmaceutically acceptable excipient, diluent,and/or carrier.

Administration of a composition or formulation can be once a day, twicea day, or more often. Frequency may be decreased during a treatmentmaintenance phase of the treatment, e.g., once every second or third dayinstead of every day or twice a day. The dose and the administrationfrequency can be adjusted based on the judgment of the treatingphysician, for example taking into account the clinical signs,pathological signs and clinical and subclinical symptoms of a disease ofthe conditions treated with the present methods, as well as thepatient's clinical history. For example, higher doses or frequency ofadministration, or a longer duration of treatment may be indicated whena patient is showing symptoms of pterygium or keloid recurrence (e.g.,blood vessel growth), or if the patient has a history of previouspterygium or keloid recurrence. Specific examples of dosage andfrequency are provided in the Examples.

It will be appreciated that the amount of an agent disclosed hereinrequired for use in treatment will vary with the route ofadministration, the nature of the condition for which treatment isrequired, and the age, body weight and condition of the patient, andwill be ultimately at the discretion of the attendant physician orveterinarian. Compositions will typically contain an effective amount ofthe active agent(s), alone or in combination. Preliminary doses can bedetermined according to animal tests, and the scaling of dosages forhuman administration can be performed according to art-acceptedpractices.

Length of treatment, i.e., number of days, will be readily determined bya physician treating the subject; however the number of days oftreatment may range from about 1 day to about 365 days. As provided bythe present methods, and discussed below, the efficacy of treatment canbe monitored during the course of treatment to determine whether thetreatment has been successful, or whether additional (or modified)treatment is necessary.

Kits

In certain embodiments, the present disclosure provides kits fortreating pterygium or keloid recurrence in a subject. The kits caninclude an anti-VEGF antibody (e.g., bevacizumab, Ranibizumab,Pegaptanib) or anti-VEGF agent (e.g., Anecortave acetate (angiostaticsteroid), VEGF-trap (a fusion protein), Squalamine lactate (smallmolecule), Combretastatin A4 Prodrug (an antitubulin/antiangiogenicagent), AdPEDF (Adenovector pigment epithelium-derived factor), Cand5(siRNA), and TG100801 (VEGF receptor blocker)), and one or more of ananti-inflammatory steroid (suitable for application to eye or skin)(e.g., triamcinolone acetonide, prednisolone acetate 1% and/ordifluprednate 0.05% (for eye), or alclometasone 0.005% cream,diflorasone 0.005% cream, fluocinonide cream 0.1%, and/or prednicarbatetopical 0.1% cream or ointment (for skin), and an NSAID (e.g.,diclofenac 1% gel, diclofenac 1.5% solution (topical)). The skilledartisan will appreciate that the dosages of the above anti-inflammatorysteroids and NSAIDs may be varied without departing from the nature ofthe present disclosure, and thus other dosages are also encompassed bythe present disclosure. The skilled artisan will know which dosages ofthe anti-inflammatory steroids and NSAIDs disclosed herein may be safelyand effectively administered to a subject according to the standard ofcare and knowledge in the art, and can include the anti-inflammatorysteroids and NSAIDs described herein.

In a specific embodiment, a kit includes bevacizumab; and one or more ofprednisolone acetate 1% and difluprednate 0.05% and triamcinoloneacetonide. In another specific embodiment, a kit includes bevacizumaband diclofenac 0.1% (and/or ketorolac tromethamine ophthalmic solution0.4%, bromfenac ophthalmic solution 0.09%, and/or nepafenac ophthalmicsuspension 0.1%). In another embodiment, the kit includes bevacizumab;one or more of: prednisolone acetate 1%, difluprednate 0.05%, andtriamcinolone acetonide; and diclofenac 0.1%. In certain embodiments,the kit is for treating pterygium. The kit can further optionallyinclude instructions for use. The kit can further optionally include(e.g., comprise, consist essentially of, consist of) syringes orapplicators preloaded with the above mentioned agents and/or vialscontaining one or more of the agents.

In another embodiment, the kit includes bevacizumab; and one or more ofalclometasone 0.005% cream, diflorasone 0.005% cream, fluocinonide cream0.1%, and prednicarbate topical 0.1% cream or ointment. In anotherembodiment, the kit includes bevacizumab; and diclofenac 1% solution. Inanother embodiment, the kit include bevacizumab; one or more ofalclometasone 0.005% cream, diflorasone 0.005% cream, fluocinonide cream0.1%, and prednicarbate topical 0.1% cream or ointment; and diclofenac1% solution. In certain embodiments, the kit is for treating keloidrecurrence. The kit can further optionally include instructions for use.The kit can further optionally include syringes or applicators preloadedwith the above mentioned agents and/or vials containing one or more ofthe agents.

The kits, regardless of type, will generally include one or morecontainers into which the biological agents (e.g., inhibitors) areplaced and, preferably, suitably aliquotted. The components of the kitsmay be packaged either in aqueous media or in lyophilized form.

In accordance with the present invention, there may be employedconventional molecular biology, microbiology, recombinant DNA,immunology, cell biology and other related techniques within the skillof the art. See, e.g., Sambrook et al., (2001) Molecular Cloning: ALaboratory Manual. 3rd ed. Cold Spring Harbor Laboratory Press: ColdSpring Harbor, N.Y; Sambrook et al., (1989) Molecular Cloning: ALaboratory Manual. 2nd ed. Cold Spring Harbor Laboratory Press: ColdSpring Harbor, N.Y; Ausubel et al., eds. (2005) Current Protocols inMolecular Biology. John Wiley and Sons, Inc.: Hoboken, N.J.; Bonifacinoet al., eds. (2005) Current Protocols in Cell Biology. John Wiley andSons, Inc.: Hoboken, N.J.; Coligan et al., eds. (2005) Current Protocolsin Immunology, John Wiley and Sons, Inc.: Hoboken, N.J.; Coico et al.,eds. (2005) Current Protocols in Microbiology, John Wiley and Sons,Inc.: Hoboken, N.J.; Coligan et al., eds. (2005) Current Protocols inProtein Science, John Wiley and Sons, Inc.: Hoboken, N.J.; Enna et al.,eds. (2005) Current Protocols in Pharmacology John Wiley and Sons, Inc.:Hoboken, N.J.; Hames et al., eds. (1999) Protein Expression: A PracticalApproach. Oxford University Press: Oxford; Freshney (2000) Culture ofAnimal Cells: A Manual of Basic Technique. 4th ed. Wiley-Liss; amongothers. The Current Protocols listed above are updated several timesevery year.

The following examples are meant to illustrate, not limit, theinvention.

Examples Materials and Methods Treatment Protocol

A topical anesthetic (proparacaine 0.5% eye drops or Alcainemanufactured by Alcon Laboratories Inc.) was applied to the eyefollowing recent pterygiectomy (from ˜ 5 days to 2 months followingsurgery). The patients were examined to identify vascular growth on eyeconjunctiva. A syringe containing bevacizumab (Avastin®)(Genentech/Roche) at a mixture of 2.5 mg/0.1 mL was injected into theconjunctiva, and the plunger was drawn back to determine if blood waspresent in the syringe hub. If blood was present (indicating presence ofblood vessel), the syringe needle was withdrawn. Bevacizumab wasinjected into areas adjacent to vascular growth, where blood vesselswere dilated and numerous. If bleeding occurred (e.g., in the form ofsubconjunctival or subcutaneous hematoma), tamponade with Q-tips® wasperformed until bleeding stopped; then injection was resumed. In somecases, patients were injected with a combination of bevacizumab andKenalog®) (KENALOG-40 (triamcinolone acetonide), suspension in an amountbetween 0.1 mL to 1 mL (1 mg to 40 mg).

Either before or after bevacizumab or bevacizumab/Kenalog® injection(time between treatments varied), patients were treated with one or bothof anti-inflammatory steroid eye drops and NSAID eye drops.Anti-inflammatory steroid drops (prednisolone acetate 1% (Allergan))were given for 6 weeks with frequency range from once a day to 4 times aday (every 6 hours) to every hour (24 drops/day). NSAID eye drops(Diclofenac 0.1% (Bausch and Lomb)) drops (1 mg/mL to 100 mg/mL) weregiven multiple times, 4 times a day every 6 hours for 3 to 6 months.

Patients were followed up to monitor for appearance of new blood vesselgrowth in the eye. The presence of new vessel growth was an indicationfor further treatment (e.g., with injection and/or eye drops (steroidand/or NSAID). In some cases, during the first surgery to removepterygium, the pterygium was only partially removed (e.g., to preventnecrosis). Thus, in some patients, additional surgeries weresubsequently performed to remove remaining quadrants of the pterygium.

Treatment of Chronic Pterygium with Bevacizumab with or withoutCombination Therapy

Five patients (Patients 1-5) underwent surgery to remove primarypterygium (pterygiectomy) from the cornea of one or both eyes followedby conjunctival grafting were treated as described below.

1. Patient 1—Right Eye

A 35 year old patient with pterygium of the right eye was treated bysurgical removal of the pterygium and conjunctival grafting. Thetimeline of the patient's treatment is shown in FIG. 1. Followingrecurrence of the pterygium, a second surgical removal of the pterygiumand conjunctival grafting procedure was performed 208 days afterperforming the first procedure. Following a further recurrence of thepterygium, a third surgical removal of the pterygium and conjunctivalgrafting procedure was performed 371 days after performing the firstprocedure.

The following drug treatments were given on the number of days indicatedafter the third surgical removal and conjunctival grafting procedure:

Day 47: Bevacizumab injection.

Day 164: Bevacizumab injection.

Day 222: Bevacizumab+Kenalog® injection.

Day 315: Bevacizumab injection.

Day 372: NSAID eye drops treatment started.

New blood vessel growth was observed at the time of the bevacizumabinjections.

No pterygium regrowth was observed as of about 520 days followingperformance of the third surgical removal and conjunctival graftingprocedure with the drug treatment as described above.

2. Patient 2—Left Eye

A 58 year old patient presented with pterygium of the left eye. Thetimeline of the patient's treatment is shown in FIG. 2. The patient wastreated by surgical removal of the pterygium and corneal regrafting.Following recurrence of the pterygium, a second surgical removal of thepterygium and conjunctival grafting procedure was performed 315 daysafter performing the first procedure.

The following drug treatments were given on the number of days indicatedafter the second surgical removal and conjunctival grafting procedure:

Day 17: Bevacizumab injection.

Following a further recurrence of the pterygium, a third surgicalremoval of the pterygium and conjunctival grafting procedure wasperformed 486 days after performing the first procedure.

The following drug treatments were given on the number of days indicatedafter the third surgical removal and conjunctival grafting procedure:

Day 14: NSAID eye drops treatment started.

Day 15: Bevacizumab and Kenalog® injection.

Day 32: Bevacizumab injection.

Day 47: Bevacizumab and Kenalog® injection.

Day 231: Bevacizumab injection.

In addition, a fourth surgical procedure, superior growth removal, wasperformed 199 days following the third pterygium removal and surgicalgrafting procedure. It is noted that superior growth is not pterygiumregrowth. The superior growth was a portion of pterygium that was notremoved in previous surgeries. Only one quadrant of the pterygium couldbe removed each time (in each surgery), because removing too manyquadrants at once could cause necrosis of the eye.

No pterygium regrowth was observed as of about 240 days followingperformance of the third surgical removal and conjunctival graftingprocedure with the drug treatment described above.

3. Patient 3

A 52 year old patient with pterygium of both the left eyes was treatedas described below. A timeline of the patient's treatment is shown inFIGS. 3A and 3B.

Left Eye

The timeline for the treatment of the patient's left eye is shown inFIG. 3A. The patient's left eye was treated by surgical removal togetherwith grafting of an amniotic membrane manufactured by IOP Ophthalmics onDay 0. The following drug treatments were given on the number of daysindicated after the surgery was performed on the left eye:

Day 53: Bevacizumab and Kenalog® injection into left eye.

No NSAID drop was used in the left eye.

Pterygium completely regrew in the patient's left eye.

Right Eye

The timeline for the treatment of the patient's right eye is shown inFIG. 3B. The patient's left eye was treated by surgical removal togetherwith grafting of an amniotic membrane manufactured by IOP Ophthalmics.This procedure was performed 189 days following surgical removal andgrafting procedure performed on the left eye. The following drugtreatments were given on the number of days indicated after surgery wasperformed on the right eye:

Day 1 Steroid eye drops treatment started.

Day 8: Bevacizumab injection into right eye.

Day 9: NSAID eye drops treatment started.

Day 28: Bevacizumab injection into right eye.

No regrowth of pterygium into the patient's right eye was observed as ofabout 130 days following performance of the surgical removal andamniotic membrane grafting procedure on the right eye.

4. Patient 4—Right Eye.

A 70 year old patient presented with pterygium of the right eye. Thetimeline of the patient's treatment is shown in FIG. 4. The patient wastreated on Day 0 by surgical pterygiectomy and conjunctival grafting.The following drug treatments were given on the number of days indicatedafter surgery was performed on the right eye:

Day 1: Steroid eye drops first used in right eye.

Day 34: Bevacizumab and Kenalog® injections into right eye.

Day 47: NSAID eye drop first used in right eye.

Day 78: Bevacizumab injection into right eye.

Day 90: Bevacizumab injection into right eye.

On day 74 following the initial surgery, a further pterygiectomyprocedure was carried out on the patient's right eye to remove pterygiumthat could not be removed during the first surgery because of scarring.

No regrowth of pterygium into the patient's right eye was observed as ofabout 250 days following performance of first surgical procedure toremove pterygium from the patient's right eye.

5. Patient 5—Right Eye

A 66 year old patient presented with pterygium of the right eye. Atimeline of the patient's treatment is shown in FIG. 5. The patient wastreated by surgical pterygiectomy and amniotic membrane grafting(manufactured by Biotissue). The following drug treatments were given onthe number of days indicated after surgery was performed on thepatient's right eye:

Day 1: Steroid eye drops and NSAID eye drops treatment started.

Day 34: Bevacizumab and Kenalog® injections into right eye.

Day 6: Bevacizumab injection into right eye.

Day 20: Bevacizumab injection into right eye.

No regrowth of pterygium into the patient's right eye was observed as ofabout 90 days following performance of surgical procedure to removepterygium from the patient's right eye.

The various treatment regimens (combination therapies) and results forpatients 1-5 are summarized in Table 4, below:

TABLE 4 Steroid (injected Topical Bevacizumab and/or NSAID PterygiumPatient Treatment eye drops) (eye drops) Recurrence 1 Right eye Yes YesYes No 2 Left eye Yes Yes Yes No 3 Right Eye Yes Yes Yes No Left Eye YesYes No Yes 4 Right Eye Yes Yes Yes No 5 Right Eye Yes Yes Yes No

The above results demonstrated that pterygium regrowth was inhibitedwhen the patient received a combination therapy including administrationof bevacizumab, an anti-inflammatory steroid (injected or topical), anda topical NSAID, and that the combination therapy was superior totreatment with bevacizumab alone.

A number of embodiments of the invention have been described.Nevertheless, it will be understood that various modifications may bemade without departing from the spirit and scope of the invention.Accordingly, other embodiments are within the scope of the followingclaims.

What is claimed is:
 1. A method for treating keloid recurrence, themethod comprising administering (1) an anti-VEGF agent to a site where akeloid has been removed from a subject.
 2. The method of claim 1,wherein the anti-VEGF agent is an antibody or small molecule inhibitorof VEGF signaling.
 3. The method of claim 1, wherein the anti-VEGF agentis the antibody bevacizumab.
 4. The method according to claim 3, whereinthe bevacizumab is administered at a dose of 10 mg.
 5. The method ofclaim 1, wherein the anti-VEGF agent is administered by injection ortopically.
 6. The method of claim 5, wherein the anti-VEGF agent isinjected at or adjacent to a site of new blood vessel growth at the siteof keloid removal.
 7. The method of claim 1, further comprisingadministering to the site (2) an anti-inflammatory steroid or (3) anon-steroidal anti-inflammatory drug (NSAID), or both.
 8. The method ofclaim 1, wherein the anti-inflammatory steroid is applied topically,systemically, or by injection to the site of keloid removal.
 9. Themethod of claim 1, wherein the anti-inflammatory steroid isalclometasone, diflorasone, fluocinonide, or prednicarbate.
 10. Themethod of claim 1, wherein the NSAID is diclofenac.
 11. The method ofclaim 1, further comprising administering to the site one or moresubsequent administrations of one or more of an anti-VEGF agent, ananti-inflammatory steroid, and an NSAID.